Effects of the arbuscular mycorrhizal fungus Glomus mosseae on growth and metal uptake by four plant species in copper mine tailings

A greenhouse experiment was conducted to evaluate the potential role of arbuscular mycorrhizal fungi (AMF) in encouraging revegetation of copper (Cu) mine tailings. Two native plant species, Coreopsis drummondii and Pteris vittata, together with a turf grass, Lolium perenne and a leguminous plant Trifolium repens associated with and without AMF Glomus mosseae were grown in Cu mine tailings to assess mycorrhizal effects on plant growth, mineral nutrition and metal uptake. Results indicated that symbiotic associations were successfully established between G. mosseae and all plants tested, and mycorrhizal colonization markedly increased plant dry matter yield except for L. perenne. The beneficial impacts of mycorrhizal colonization on plant growth could be largely explained by both improved P nutrition and decreased shoot Cu, As and Cd concentrations. The experiment provided evidence for the potential use of local plant species in combination with AMF for ecological restoration of metalliferous mine tailings.     

Arbuscular mycorrhizal fungi can alleviate the adverse effects of chlorothalonil on Oryza sativa L

A glasshouse pot experiment was conducted to investigate the effect of the fungicide chlorothalonil on the growth of upland rice, in the absence or presence of the arbuscular mycorrhizal fungus (AMF) Glomus mosseae (NM and GM treatments). The plants were grown with three concentrations of chlorothalonil (0, 50 and 100 mg kg(-1) soil). Mycorrhizal colonization decreased significantly with increasing chlorothalonil concentrations. Plant biomass decreases were smaller in GM plants than in non-mycorrhizal (NM) plants. Mycorrhizal dependency was the highest with 50 mg kg(-1) chlorothalonil. Chlorothalonil affected physiological processes in upland rice irrespective of inoculation. Chlorothalonil at 50 and 100 mg kg(-1) increased ascorbate peroxidase (APX) activity and soluble protein concentrations in shoots and roots of NM upland rice. However, values of APX, catalase (CAT) and peroxidase (POD) were reduced more in GM plants than in NM plants. These results showed that chlorothalonil induced oxidative stress in upland rice and it is needed to evaluate the side effects of chlorothalonil on rice and AMF.     

Arbuscular mycorrhizal fungi can decrease the uptake of uranium by subterranean clover grown at high levels of uranium in soil

Subterranean clover inoculated or not with the arbuscular mycorrhizal (AM) fungus Glomus intraradices was grown on soil containing six levels of 238U in the range 0-87 mg kg(-1). Increasing U concentration in soil enhanced the U concentration in roots and shoots of both mycorrhizal and nonmycorrhizal plants but had no significant effects on plant dry matter production or root AM colonization. Mycorrhizas increased the shoot dry matter and P concentration in roots and shoots, while in most cases, it decreased the Ca, Mg and K concentrations in plants. The AM fungus influenced U concentration in plants only in the treatment receiving 87 mg U kg(-1) soil. In this case, U concentration in shoots of nonmycorrhizal plants was 1.7 times that of shoots of mycorrhizal plants. These results suggested that mycorrhizal fungi can limit U accumulation by plants exposed to high levels of U in soil.     

Arbuscular mycorrhizae increase the arsenic translocation factor in the As hyperaccumulating fern Pteris vittata L

Phytoremediation techniques are receiving more attention as decontaminating strategies. Phytoextraction makes use of plants to transfer contaminants from soil to the aboveground biomass. This research is devoted to study the effects of arbuscular mycorrhizae (AM) on growth and As hyperaccumulation in the Chinese brake fern Pteris vittata. We grew for 45 days P. vittata sporophytes, infected or not infected with the AM fungi Glomus mosseae or Gigaspora margarita, in a hydroponic system on quartz sand. As-treated plants were weekly fed with 25 ppm As. The As treatment produced a dramatic increase of As concentration in pinnae and a much lower increase in roots of both mycorrhizal and control plants. Mycorrhization increased pinnae dry weight (DW) (G. margarita = G. mosseae) and leaf area (G. margarita > G. mosseae), strongly reduced root As concentration (G. mosseae > G. margarita), and increased the As translocation factor (G. mosseae > G. margarita). The concentration of phosphorus in pinnae and roots was enhanced by both fungi (G. margarita > G. mosseae). The quantitatively different effects of the two AM fungi on plant growth as well as on As and P distribution in the fern suggest that the As hyperaccumulation in P. vittata can be optimized by a careful choice of the symbiont.     

Effects of arbuscular mycorrhizal inoculation on plants growing on arsenic contaminated soil

Arbuscular mycorrhizal fungi (AMF) may play an important role in phytoremediation of As-contaminated soil. In this study the effects of AMF (Glomus mosseae, Glomus intraradices and Glomus etunicatum) on biomass production and arsenic accumulation in Pityrogramma calomelanos, Tagetes erecta and Melastoma malabathricum were investigated. Soil (243 +/- 13 microg As g(-1)) collected from Ron Phibun District, an As-contaminated area in Thailand, was used in a greenhouse experiment. The results showed different effects of AMF on phytoremediation of As-contaminated soil by different plant species. For P. calomelanos and T. erecta, AMF reduced only arsenic accumulation in plants but had no significant effect on plant growth. In contrast, AMF improved growth and arsenic accumulation in M. malabathricum. These findings show the importance of understanding different interactions between AMF and their host plants for enhancing phytoremediation of As-contaminated soils.     

Zinc accumulation in Solanum nigrum is enhanced by different arbuscular mycorrhizal fungi

Solanum nigrum was found to proliferate in sediments with high levels of metal pollution. The effect of Zn on plant growth and tissue metal accumulation was assessed. The response of the plant to the inoculation with four different isolates of arbuscular mycorrhizal fungi (AMF) (Glomus sp. BEG140, Glomus claroideum, Glomus mosseae and Glomus intraradices) was studied. While the isolates of AMF did not have a significant (P<0.05) influence on mycorrhizal colonisation, increasing Zn concentration to high levels (500 and 1000 mgkg(-1)) induced significant (P<0.05) decrease of the AMF colonisation. In general, the presence of AMF did not affect the growth and biomass of S. nigrum individuals. However, the level of metal in the matrix affected S. nigrum growth; plants grown at 100 mgkg(-1) had significantly (P<0.05) lower leaf, stem, root and total biomass than control ones and plants growing at 500 and 1000 mgkg(-1) had the significantly (P<0.05) lowest biomass. Plants inoculated with the AMF G. claroideum and G. intraradices presented significantly (P<0.05) higher Zn accumulation in all plant tissues. In general, the stem tissues had the higher Zn content while the leaves registered the lowest values, which indicate a high translocation of the metal. AMF inoculation had no significant (P<0.05) influence on the metal translocation within the plant. This study suggests that inoculation with the AMF G. claroideum or G. intraradices, can enhance the Zn accumulation in the tissues of S. nigrum, not affecting the plant translocation capacities.     

Arbuscular mycorrhizal fungi mediated uptake of lanthanum in Chinese milk vetch (Astragalus sinicus L.)

A glasshouse experiment was carried out to study the effect of mycorrhizal formation by Gigaspora margarita, Glomus intraradices or Acaulospora laevis on plant growth and lanthanum (La) uptake of Astragalus sinicus L. in soils spiked with La at five levels (0, 1, 5, 10 and 20 mg kg(-1)). La application decreased mycorrhizal infection frequency and activity of fungal succinate dehydrogenase and alkaline phosphatase. Increasing La concentrations in soil led to increased La accumulation in tissues of both mycorrhizal and non-mycorrhizal plants, but inoculation with Gig. margarita or G. intraradices reduced La concentrations in shoots and roots at higher concentrations of La in the soil. Plant biomass and P concentrations in shoots and roots were reduced by La application, but increased by inoculation with Gig. margarita or G. intraradices. The results showed that appropriate AM fungi colonization could be effective in alleviating La toxicity in A. sinicus.     

Arbuscular mycorrhizal fungi differentially affect the response to high zinc concentrations of two registered poplar clones

The effects of a high concentration of zinc on two registered clones of poplar (Populus alba Villafranca and Populus nigra Jean Pourtet), inoculated or not with two arbuscular mycorrhizal fungi (Glomus mosseae or Glomus intraradices) before transplanting them into polluted soil, were investigated, with special regard to the extent of root colonization by the fungi, plant growth, metal accumulation in the different plant organs, and leaf polyamine concentration. Zinc accumulation was lower in Jean Pourtet than in Villafranca poplars, and it was mainly translocated to the leaves; the metal inhibited mycorrhizal colonization, compromised plant growth, and, in Villafranca, altered the putrescine profile in the leaves. Most of these effects were reversed or reduced in plants pre-inoculated with G. mosseae. Results indicate that poplars are suitable for phytoremediation purposes, confirming that mycorrhizal fungi can be useful for phytoremediation, and underscore the importance of appropriate combinations of plant genotypes and fungal symbionts.     

Cu污染土壤接种丛枝菌根真菌对旱稻生长的影响

采用盆栽实验的方法,研究了不同Cu处理水平(0、100和200 mg/kg)下,接种丛枝菌根真菌(arbuscular my-corrhizal fungi,AMF)Glomus mosseae对旱稻(Oryzal Sativa L.)生长的影响。结果表明,未添加Cu处理下,旱稻菌根侵染率可达69%,随着土壤中Cu添加量增加,旱稻菌根侵染率显著下降(P<0.05)。与未接种处理相比,接种处理显著提高100mg/kg Cu处理下根系生物量以及200 mg/kg Cu处理下地上部分生物量(P<0.05);接种处理显著降低了100 mg/kg Cu处理下旱稻地下部Cu含量,却显著增加了200 mg/kg Cu处理下旱稻地上部分以及地下部分Cu含量(P<0.05)。     

Phytoextraction by arsenic hyperaccumulator Pteris vittata L. from six arsenic-contaminated soils: Repeated harvests and arsenic redistribution

This greenhouse experiment evaluated arsenic removal by Pteris vittata and its effects on arsenic redistribution in soils. P. vittata grew in six arsenic-contaminated soils and its fronds were harvested and analyzed for arsenic in October, 2003, April, 2004, and October, 2004. The soil arsenic was separated into five fractions via sequential extraction. The ferns grew well and took up arsenic from all soils. Fern biomass ranged from 24.8 to 33.5 g plant(-1) after 4 months of growth but was reduced in the subsequent harvests. The frond arsenic concentrations ranged from 66 to 6,151 mg kg(-1), 110 to 3,056 mg kg(-1), and 162 to 2,139 mg kg(-1) from the first, second and third harvest, respectively. P. vittata reduced soil arsenic by 6.4-13% after three harvests. Arsenic in the soils was primarily associated with amorphous hydrous oxides (40-59%), which contributed the most to arsenic taken up by P. vittata (45-72%). It is possible to use P. vittata to remediate arsenic-contaminated soils by repeatedly harvesting its fronds.     

Effect of arbuscular mycorrhizal fungus (Glomus caledonium) on the accumulation and metabolism of atrazine in maize (Zea mays L.) and atrazine dissipation in soil

Effects of an arbuscular mycorrhizal (AM) fungus (Glomus caledonium) on accumulation and metabolism of atrazine in maize grown in soil contaminated with different concentrations of atrazine were investigated in a series of pot experiments. Roots of mycorrhizal plants accumulated more atrazine than non-mycorrhizal roots. In contrast, atrazine accumulation in shoot decreased in mycorrhizal compared with non-mycorrhizal plants. No atrazine derivatives were detected in the soil, either with or without mycorrhizal colonization. However, atrazine metabolites, deethylatrazine (DEA) and deisopropylatrazine (DIA), were detected in plant roots and the AM colonization enhanced the metabolism. After plant harvest atrazine concentrations decreased markedly in the soils compared to the initial concentrations. The decreases were the most in rhizosphere soil and then near-rhizosphere soil and the least in bulk soil. Mycorrhizal treatment enhanced atrazine dissipation in the near-rhizosphere and bulk soils irrespective of atrazine application rates.     

Survival strategies of plants associated with arbuscular mycorrhizal fungi on toxic mine tailings

A field survey of metal concentrations and arbuscular mycorrhizal (AM) components of plants growing on five mining sites was conducted in Chenzhou City, Hunan Province, Southern China and a control site in Hong Kong. Significant differences were observed in the average concentrations of total heavy metals (Pb, Zn, Cu, Cd) and one metalloid (As) in contaminated soils compared with the control site. Gramineae and Compositae were the dominant plant families growing on mine tailings, with Chrysanthemum moritolium (common chrysanthemum), Cynodon dactylon (Bermuda grass), Miscanthus florodulus (Sword grass) and Pteris vittata (Ladder brake fern) commonly found at all sites. AM fungal colonization was detected in most of the plants. Comparing the four common plant species, three components of mycorrhizal colonization (arbuscules, vesicles and coiled hyphae) were found in the roots of C. dactylon and P. vittata growing at Do Shun Long (DSL) mine site. Concentrations of As in fronds were 24-fold higher than in roots of P. vittata with the highest mycorrhizal colonization rate (73%) among all sampling sites. Extensive mycorrhizal colonization (85%) was also recorded in the roots of C. dactylon with As accumulation 57 times higher than in shoots. The four common plants found in metal contaminated sites had developed different strategies for survival in the contaminated sites with the aid of indigenous AM fungi.     

Solanum nigrum grown in contaminated soil: effect of arbuscular mycorrhizal fungi on zinc accumulation and histolocalisation

Zn tissue accumulation in Solanum nigrum grown in a non-contaminated and a naturally contaminated Zn matrix and the effect of inoculation with different arbuscular mycorrhizal fungi (AMF) on metal uptake were assessed. S. nigrum grown in the contaminated soil always presented higher Zn accumulation in the tissues, accumulating up to 1622 mg Zn kg(-1). The presence of both Glomus claroideum and Glomus intraradices enhanced the uptake and accumulation of Zn by S. nigrum (up to 83 and 49% higher Zn accumulation, respectively). The main deposits of the metal were found in the intercellular spaces and in the cell walls of the root tissues, as revealed by autometallography, with the inoculation with different AMF species causing no differences in the location of Zn accumulation. These findings indicate that S. nigrum inoculated with selected heavy metal tolerant AMF presents extracting and accumulating capacities, constituting a potentially suitable remediation method for Zn polluted soils.     

Arsenic hyperaccumulation by Pteris vittata from arsenic contaminated soils and the effect of liming and phosphate fertilisation

Pot experiments were carried out to investigate the potential of phytoremediation with the arsenic hyperaccumulator Pteris vittata in a range of soils contaminated with As and other heavy metals, and the influence of phosphate and lime additions on As hyperaccumulation by P. vittata. The fern was grown in 5 soils collected from Cornwall (England) containing 67-4550 mg As kg(-1) and different levels of metals. All soils showed a similar distribution pattern of As in different fractions in a sequential extraction, with more than 60% of the total As being associated with the fraction thought to represent amorphous and poorly-crystalline hydrous oxides of Fe and Al. The concentration of As in the fronds ranged from 84 to 3600 mg kg(-1), with 0.9-3.1% of the total soil As being taken up by P. vittata. In one soil which contained 5500 mg Cu kg(-1) and 1242 mg Zn kg(-1), P. vittata suffered from phytotoxicity and accumulated little As (0.002% of total). In a separate experiment, neither phosphate addition (50mg P kg(-1) soil) nor liming (4.6 g CaCO3 kg(-1) soil) was found to affect the As concentration in the fronds of P. vittata, even though phosphate addition increased the As concentration in the soil pore water. Between 4 and 7% of the total soil As was taken up by P. vittata in 4 cuttings in this experiment. The results indicate that P. vittata can hyperaccumulate As from naturally contaminated soils, but may be suitable for phytoremediation only in the moderately contaminated soils.     

The role of arbuscular mycorrhiza in zinc uptake by red clover growing in a calcareous soil spiked with various quantities of zinc

Three pot experiments were conducted to investigate the role of arbuscular mycorrhiza (AM) in Zn uptake by red clover. Plants inoculated with Glomus mosseae and uninoculated controls were grown in a sterile calcareous soil in 'Plexiglas' (Acrylic) containers with two nylon net partitions (30 microm mesh) to separate the central root zone from the two outer hyphal zones. The effects of mycorrhiza on plant growth and Zn uptake changed dramatically with increasing Zn addition level (range 0-1200 mg kg(-1)) in the root zone. With Zn addition levels <300 mg kg(-1), added Zn did not affect plant yield and above the critical level plant yield gradually decreased but was always higher for mycorrhizal than for controls. Below the critical Zn application rate (50 mg kg(-1)), Zn uptake was enhanced while above this level Zn translocation to the shoots decreased. At all Zn addition levels, mycorrhizal colonization increased Zn absorption and accumulation in the roots, and this may help to explain the alleviation of Zn toxicity at high Zn application rates. As expected, AM colonization enhanced P nutrition and hence yield at all added Zn levels studied. Efforts were made to obtain direct evidence for hyphal contribution to Zn uptake by applying both Zn to the hyphal growth zone and additional P to the root zone to avoid the 'growth dilution effect'. The data demonstrate that mycorrhizal hyphae could absorb Zn directly from the soil and then transfer it to the plant roots. The hyphal contribution to Zn uptake by the host plant reached its maximum value at the Zn addition level of 50 mg kg(-1), in which Zn uptake via the extramatrical hyphae comprised 22% of total uptake, thus confirming the critical Zn application level found previously.     

Interactions between arbuscular mycorrhizae and heavy metals under sand culture experiment

A sand culture experiment was established to determine interactions between arbuscular mycorrhizae and heavy metals. Mycorrhizal infection rates, spore densities, maize root and shoot weights, and heavy metal contents in maize were as indexes of responses of arbuscular mycorrhizal fungi (Acaulospora laevis, Glomus caledonium and Glomus manihotis) to heavy metals (Cu and Cd). The mycorrhizal infection rates of G. caledonium were the highest among these three mycorrhizal fungi, but the sporulating ability of G. caledonium was the poorest in the heavy metal treatments. The shoot and root weights of non-mycorrhizal plants were usually greater than those of mycorrhizal plants when the Cu concentrations in solutions are less than 3 mg l(-1) or Cd concentrations less than 1 mg l(-1). When Cd concentrations were 0.5 and 1 mg(-1), the root and shoot weights of plants inoculated with A. laevis were significantly (p < 0.05) lower than those of other treatments. Copper concentrations in shoots of mycorrhizal plants were higher than those of non-mycorrhizal ones at all Cu concentrations in solution, especially at low Cu concentrations. As to A. laevis, Cu concentrations in roots and shoots of the host were higher than those of non-mycorrhizal plants in these treatments. Thus A. laevis was sensitive to Cu and Cd, especially Cd, and G. caledonium was more tolerant to these two heavy metals. It is suggested that G. caledonium might be a promising mycorrhizal fungus for bioremediation of heavy metal contaminated soil.     

Effects of arsenic on concentration and distribution of nutrients in the fronds of the arsenic hyperaccumulator Pteris vittata L

Pteris vittata was the first terrestrial plant known to hyperaccumulate arsenic (As). However, it is unclear how As hyperaccumulation influences nutrient uptake by this plant. P. vittata fern was grown in soil spiked with 0-500 mg As kg(-1) in the greenhouse for 24 weeks. The concentrations of essential macro- (P, K, Ca, and Mg) and micro- (Fe, Mn, Cu, Zn, B and Mo) elements in the fronds of different age were examined. Both macro- and micronutrients in the fronds were found to be within the normal concentration ranges for non-hyperaccumulators. However, As hyperaccumulation did influence the elemental distribution among fronds of different age of P. vittata. Arsenic-induced P and K enhancements in the fronds contributed to the As-induced growth stimulation at low As levels. The frond P/As molar ratios of 1.0 can be used as the threshold value for normal growth of P. vittata. Potassium may function as a counter-cation for As in the fronds as shown by the As-induced K increases in the fronds. The present findings not only demonstrate that P. vittata has the ability to maintain adequate concentrations of essential nutrients while hyperaccumulating As from the soil, but also have implications for soil management (fertilization in particular) of P. vittata in As phytoextraction practice.     

EDDS and EDTA-enhanced zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi grown in contaminated soil

The effect of two different chelating agents [EDTA and EDDS (S,S-ethylenediaminedissucinic acid)] on Zn tissue accumulation in Solanum nigrum L. grown in a naturally contaminated soil was assessed. Under those conditions, the response of the plant to the inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF)--Glomus claroideum and Glomus intraradices--was also studied. Plants grown in the local contaminated soil (Zn levels of 433mg kg(-1)) accumulated up to 1191mg kg(-1) of Zn in the roots, 3747mg kg(-1) in the stems and 3409mg kg(-1) in the leaves. S. nigrum plants grown in the same soil spiked with extra Zn (Zn levels of 964mg kg(-1)) accumulated up to 4735, 8267 and 7948mg Zn kg(-1) in the leaves, stems and roots, respectively. The addition of EDTA promoted an increase in the concentration of Zn accumulated by S. nigrum of up to 231% in the leaves, 93% in the stems and 81% in the roots, while EDDS application enhanced the accumulation in leaves, stems and roots up to 140, 124 and 104%, respectively. In the stems, the presence of Zn was predominantly detected in the cortex collenchyma cells, the starch sheath and the internal phloem and xylem parenchyma, and the addition of chelating agents did not seem to have an effect on the localisation of accumulation sites. The devise of a chelate-enhanced phytoextraction strategy, using chelating agents and AMF, is discussed.     

Arsenic accumulation by two brake ferns growing on an arsenic mine and their potential in phytoremediation

In an area near an arsenic mine in Hunan Province of south China, soils were often found with elevated arsenic levels. A field survey was conducted to determine arsenic accumulation in 8 Cretan brake ferns (Pteris cretica) and 16 Chinese brake ferns (Pteris vittata) growing on these soils. Three factors were evaluated: arsenic concentration in above ground parts (fronds), arsenic bioaccumulation factor (BF; ratio of arsenic in fronds to soil) and arsenic translocation factor (TF; ratio of arsenic in fronds to roots). Arsenic concentrations in the fronds of Chinese brake fern were 3-704 mg kg-1, the BFs were 0.06-7.43 and the TFs were 0.17-3.98, while those in Cretan brake fern were 149-694 mg kg-1, 1.34-6.62 and 1.00-2.61, respectively. Our survey showed that both ferns were capable of arsenic accumulation under field conditions. With most of the arsenic being accumulated in the fronds, these ferns have potential for use in phytoremediation of arsenic contaminated soils.     

Application of manure and compost to contaminated soils and its effect on zinc accumulation by Solanum nigrum inoculated with arbuscular mycorrhizal fungi

Zn accumulation in Solanum nigrum grown in naturally contaminated soil in the presence of different types of organic amendments was assessed. Under the same conditions, the response of the plant to inoculation with two different isolates of arbuscular mycorrhizal fungi (AMF) (Glomus claroideum and Glomus intraradices) was also evaluated. S. nigrum grown in the non-amended soil always presented higher Zn accumulation in the tissues, with the addition of amendments inducing reductions of up to 80 and 40%, for manure and compost, respectively, and enhancing plant biomass yields. The establishment of S. nigrum in the Zn contaminated soil combined with the application of amendments led to a 70-80% reduction in the amount of Zn leached through the soil. The use of S. nigrum in combination with manure appeared as an effective method for reducing the effects of soil contamination, diminishing Zn transfer to other environmental compartments via percolation.